JP5393264B2 - Objective optical system position adjustment device and observation device - Google Patents

Description

  The present invention relates to an objective optical system position adjusting device and an observation device.

  In recent years, it has become important to observe the inside of a biological sample such as a small animal, for example, the state of a deep brain or an organ in a living state (in vivo). Therefore, there are known an objective optical system and a microscope system having a small-diameter tip portion that can reduce the invasion to a living body and enable temporal observation in the living body for a relatively long period of time (for example, Patent Document 1 and Patent). Reference 2).

  In addition, when observing the inside of a living body over time, if an attempt is made to cause a small animal or the like to perform the same activity as normal during the observation period, the objective optical system positioned at the observation site is temporarily removed and the same is again performed during observation. It is necessary to position the objective optical system at the observation site. At this time, there is known a fixing device that positions the objective optical system with high reproducibility by inserting the objective optical system into the insertion hole of the support member fixed to the living body (see, for example, Patent Document 3). .

JP 2006-119300 A JP 2005-241671 A Japanese Patent Laying-Open No. 2005-253836

  However, in order to reduce the invasion of the living body by the support member when the objective optical system having a small-diameter tip and the fixing device are used together to minimize the invasion of the living body and observe the inside of the living body for a relatively long period of time. In addition, it is desirable to keep the inner diameter of the insertion hole slightly larger than the outer diameter of the small-diameter tip.

  Therefore, if the insertion hole is made thinner, there is only a small gap between the small diameter tip and the insertion hole when the small diameter tip removed from the insertion hole is inserted again into the insertion hole. There is a problem that alignment of the system becomes difficult. In addition, if the position of the small-diameter tip is displaced from the insertion hole and the small-diameter tip collides with the support member, a large force is applied and these components are damaged or the living body is affected by the impact at the time of the collision. There is a problem of fear.

  The present invention has been made in view of the above-described circumstances, and even if the insertion hole of the support member has a small diameter, the objective optical system is protected from damage to the objective optical system and the support member and the influence on the living body. An object of the present invention is to provide an objective optical system position adjusting device and an observation device that can easily insert a small-diameter tip portion into an insertion hole.

In order to achieve the above object, the present invention provides the following means.
The present invention relates to a position adjusting unit disposed between an objective optical system having a small-diameter tip and an observation optical system for observing light condensed by the objective optical system, and one end fixed to a living body. A substantially cylindrical support unit that removably supports the tip of the small diameter inside, the position adjusting unit holding the objective optical system, and the holding unit as the objective optical A moving mechanism that movably supports in a direction crossing the optical axis direction of the system, and when the small-diameter tip portion is inserted into and removed from the support unit, the holding portion is moved to the optical axis direction by the moving mechanism. by smoothly moved in a direction intersecting the position of the objective optical system to adjust relative to the support unit, the support unit, the other end portion, tapered progressively diameter increases toward the other end Objective optical system with inner surface To provide a device.

According to the present invention, by inserting the small-diameter tip of the objective optical system into the support unit fixed to the living body, the position of the objective optical system is positioned at the same position with respect to the living body, and a predetermined observation site Can be observed over time.
In this case, when the small diameter tip is inserted into the support unit, if the position of the small diameter tip is displaced with respect to the support unit, the tip of the small diameter tip contacts the tapered inner surface, The holding portion and the objective optical system are moved by the moving mechanism in accordance with the shape of the inner surface, so that the small-diameter tip is guided into the support unit while the position of the objective optical system is adjusted.

  This eliminates the need for high alignment accuracy for the small diameter tip, and supports the small diameter tip easily even if the inner diameter of the insertion hole is reduced to approximately the same as the outer diameter of the small diameter tip. Can be inserted into the unit. Even if the tip of the small-diameter tip comes into contact with the support unit, the impact at the time of contact is buffered by moving the objective optical system in the direction of escaping from the contact, causing damage to the objective optical system and the support unit and to the living body. The effect can be prevented.

In the above invention, the moving mechanism may include two linear guides for guiding the holding unit in directions perpendicular to each other in a plane substantially perpendicular to the optical axis direction of the objective optical system.
By doing so, the objective optical system can be smoothly moved in a plane substantially perpendicular to the optical axis direction.

Moreover, in the said invention, the said linear guide is good also as providing a bearing mechanism.
By doing in this way, an objective optical system can be moved to lubrication.

Moreover, in the said invention, the said position adjustment unit is good also as providing the control means which controls the movement of the said holding | maintenance part.
By doing so, it is possible to observe the image while stabilizing the field of view by preventing the movement of the objective optical system by restricting the movement of the holding part in a state where the small-diameter tip is inserted into the insertion hole.

In the above invention, the position adjustment unit adjusts the position of the optical axis of the objective optical system held by the holding unit so as to coincide with the position of the optical axis of the observation optical system. Means may be provided.
In this way, the position of the optical axis of the objective optical system can be easily matched with the optical axis of the observation optical system.
Moreover, this invention provides an observation apparatus provided with the objective optical system position adjustment apparatus in any one of the said.

  According to the present invention, even if the insertion hole of the support member has a small diameter, the objective optical system can be easily inserted into the insertion hole while preventing damage to the objective optical system and the support member and the influence on the living body. There is an effect.

1 is an overall configuration diagram of an objective optical system position adjusting device and an observation device according to an embodiment of the present invention. It is a whole block diagram which shows the position adjustment unit of the objective optical system position adjustment apparatus of FIG. It is a front view of the microscope attachment part of the position adjustment unit of FIG. It is a front view of the intermediate part of the position adjustment unit of FIG. It is a front view of the objective attachment part of the position adjustment unit of FIG. It is side surface sectional drawing which shows the support unit of the objective optical system position adjustment apparatus of FIG. It is a figure explaining the usage method of the objective optical system position adjustment apparatus of FIG. 1, and has shown the state which an objective optical system can move freely. It is a figure explaining the usage method of the objective optical system position adjustment apparatus of FIG. 1, and has shown the state by which the optical axis of the objective optical system was adjusted. It is a figure explaining the usage method of the objective optical system position adjustment apparatus of FIG. 1, and has shown the state when bringing an objective optical system close to a sample. It is a figure which shows the process in which an objective optical system is inserted in an insertion hole using the objective optical system position adjustment apparatus of FIG.

An objective optical system position adjusting device 1 and an observation device 100 according to an embodiment of the present invention will be described below with reference to FIGS. In the present embodiment, the case where the inside of the brain of a living mouse (living body) A is observed will be described as an example.
As shown in FIG. 1, the observation apparatus 100 according to the present embodiment is based on an upright optical microscope. A microscope (observation optical system) 2 and an objective optical system 3 attached to the microscope 2 are attached. The objective optical system position adjusting device 1 according to this embodiment including the position adjusting unit 4 and the support unit 5 fixed to the mouse A is provided.

  As the microscope 2, a general optical microscope or a laser scanning microscope is used. The microscope 2 has a unit attachment portion 6 to which the position adjustment unit 4 is attached. The unit mounting portion 6 has a structure generally used for mounting an objective optical system, and is provided with, for example, an RMS screw. By moving the unit mounting portion 6 up and down along the optical axis direction, the position in the height direction of the position adjusting unit 4 and the objective optical system 3 mounted on the unit mounting portion 6 is adjusted. . The mouse A is placed on a stage 7 movable in a horizontal plane, and the head is fixed to the stage 7 by a fixture 8.

  The objective optical system 3 has a small-diameter tip 3a. The objective optical system 3 is designed at infinity, and the light collected from the tip of the objective optical system 3 is transmitted as a substantially parallel light beam to the rear optical system.

  As shown in FIG. 2, the position adjustment unit 4 includes a microscope attachment portion 9, an intermediate portion 10, and an objective attachment portion (holding portion) 11 in order from the microscope 2 side. An X-axis guide 12a (moving mechanism, linear guide) is disposed between the microscope mounting portion 9 and the intermediate portion 10, and a Y-axis guide 12b (moving mechanism) is interposed between the intermediate portion 10 and the objective mounting portion 11. , A straight guide) is arranged. The X-axis guide 12a and the Y-axis guide 12b are movable in a direction perpendicular to each other in a plane perpendicular to the optical axis of the microscope 2, and have excellent lubricity, for example, a bearing (bearing mechanism). Is used. Thereby, the objective attachment part 11 is supported by the microscope attachment part 9 so as to be movable in a horizontal plane.

  As shown in FIG. 3, the microscope attachment portion 9 has a microscope attachment screw 9 a that can be attached to and detached from the unit attachment portion 6 of the microscope 2, and the position adjustment is performed by attaching the microscope attachment screw 9 a to the unit attachment portion 6. The unit 4 is mounted on the microscope 2. In addition, a light shielding tube 9 b extending to the vicinity of the objective mounting portion 11 is provided in the approximate center of the microscope mounting portion 9. Thereby, the optical path between the microscope 2 and the objective optical system 3 is shielded from the outside, and it is prevented that extraneous light is mixed into the irradiation light 2 from the microscope 2 and the observation light collected by the objective optical system 3. It has become so.

As shown in FIG. 4, the intermediate portion 10 is provided with an intermediate portion hole 10 a through which the light shielding tube 9 b passes substantially at the center. The intermediate hole 10a has an inner diameter dimension larger than the outer diameter dimension of the light shielding cylinder 9b so that the movement of the intermediate section 10 is not restricted by the light shielding cylinder 9b when the intermediate section 10 moves in the horizontal direction with respect to the microscope mounting portion 9. doing.
As shown in FIG. 5, the objective attachment portion 11 has an objective attachment screw hole 11a through which the rear portion of the objective optical system 3 can be attached and removed substantially through the center.

  A centering screw (optical axis adjusting means) 13 is inserted into the microscope mounting portion 9 and the objective mounting portion 11 at a position that coincides when the light shielding tube 9b and the objective mounting screw hole 11a are arranged on a substantially concentric axis. The microscope-side centering screw hole (optical axis adjusting means) 14a and the objective-side centering hole (optical axis adjusting means) 14b penetrate through in the thickness direction. The centering screw 13 has a rear portion screw portion 13a fitted in the microscope side centering screw hole 14a, and a conical portion 13b formed in a gradually tapered shape at the tip portion is inserted into the objective side centering hole 14b. Yes.

  As a result, when the rotation screw 13c provided in the middle position of the centering screw 13 is rotated in the tightening direction, the position of the objective mounting portion 11 is set to a position where the optical axis of the objective optical system 3 and the optical axis of the microscope 2 coincide with each other. Has been adjusted. In addition, notches 10b are provided at the four corners of the intermediate portion 10, whereby the intermediate portion 10 and the centering screw 13 are movable without being restricted in position relative to each other.

  Further, the microscope mounting portion 9 is provided with a fixing screw hole (restricting means) 15b into which a fixing screw (restricting means) 15a is fitted, and the fixing screw 15a is tightened so that the space between the fixing screw 15a and the microscope mounting portion 9 is increased. By fixing the shim 16 arranged on the microscope mounting portion 9 to the microscope mounting portion 9, the objective mounting portion 11 is fixed to the microscope mounting portion 9 via a connecting member 17 that connects the shim 16 and the objective mounting portion 11. It has become.

  As shown in FIG. 6, the support unit 5 has a substantially cylindrical shape having a small diameter portion 5a inserted into a living body and a large diameter portion 5b having a larger diameter than the small diameter portion 5a. An insertion hole 5c having an inner diameter that is slightly larger than the outer diameter of the small-diameter tip 3a of the objective optical system 3 extends along the longitudinal direction in the small diameter 5a. A window 5d made of a transparent material such as a cover glass is provided in the central portion that is an optical path between the optical system 3 and the sample surface.

The distal end of the small diameter portion 5a is closed by a transparent member 5e such as glass with a suitable distance between the bottom surface of the insertion hole 5c and the sample surface. Accordingly, when the small-diameter tip portion 3a is inserted into the insertion hole 5c until the tip surface thereof abuts against the bottom surface of the insertion hole 5c, the objective optical system 3 is properly focused on the sample surface. .
The large-diameter portion 5b has an inner surface 5f that continuously increases from the inner surface of the insertion hole 5c in a conical shape.

The usage method and operation of the objective optical system position adjusting apparatus 1 and the observation apparatus 100 configured as described above will be described below with reference to FIGS. 7 to 9, a part of the configuration is omitted for easy understanding of the configuration of the explanation part.
In order to observe the inside of the brain of the mouse A in vivo using the observation apparatus 100 according to the present embodiment, the support unit 5 is bonded and fixed in a state where the small-diameter portion 5a is inserted into the through-hole drilled in the skull of the mouse A. Then, the head of the mouse A is fixed to the stage 7.

First, in a state where the fixing screw 15a is loosened so that the objective optical system 3 can be moved in a substantially horizontal plane (see FIG. 7), the centering screw 13 is tightened to change the optical axis of the objective optical system 3 to the optical axis of the microscope 2. And the objective optical system 3 is fixed to the microscope 2 by tightening the fixing screw 15a (see FIG. 8). When the centering screw 13 is loosened in this state, the objective optical system 3 is held at a position where its optical axis coincides with the optical axis of the microscope 2.

Next, when the stage 7 is moved so that the insertion hole 5c is disposed substantially vertically below the objective optical system 3 to align the objective optical system 3 with the observation site, the fixing screw 15a is loosened and the objective optical system is moved. In a state in which 3 is movable (see FIG. 9), the objective optical system 3 is slowly moved downward to insert the small-diameter tip 3a into the insertion hole 5c.

At this time, if the position of the objective optical system 3 is deviated from the insertion hole 5c, the tip of the small-diameter tip 3a comes into contact with the tapered inner surface 5f (see FIG. 10), and the tip is narrow along the inner surface 5f. The diameter tip 3a is guided to the position of the insertion hole 5c while moving smoothly, and is inserted into the insertion hole 5c as it is. Once inserted to a position abuts the narrow-diameter end portion 3a to the bottom surface of the insertion hole 5c, and tighten the fixing screw 1 5a to fix the position of the objective optical system 3, to observe the brain of the mouse A in vivo by microscope 2 be able to.

  As described above, according to the present embodiment, the support unit 5 is provided with the inner surface 5f that gradually decreases in size toward the insertion hole 5c, and the objective optical system 3 can be smoothly moved in the direction intersecting the insertion direction. In this state, it is inserted into the insertion hole 5c. Thereby, even if the position of the small-diameter tip 3a is displaced from the insertion hole 5c, the position is adjusted in the process of insertion. Therefore, even if the inner diameter dimension of the insertion hole 5c is reduced to substantially the same as the outer diameter dimension of the thin tip portion 3a, the small tip portion 3a is not required to control high positional accuracy of the objective optical system 3. There is an advantage that it can be easily inserted into the insertion hole 5c.

  In addition, by reducing the diameter of the insertion hole 5c as much as possible in this way, the body of the mouse A can be observed for a relatively long period of time while further reducing the invasion of the mouse A by the support unit 5. There is an advantage that you can. Even if the tip of the small-diameter tip 3a contacts the tapered inner surface 5f, the small-diameter tip 3a and the support are supported by the smooth movement of the small-diameter tip 3a in the direction of buffering the impact caused by the contact. There is an advantage that the unit 5 can be prevented from being damaged and the influence on the living body and the observation site due to the impact at the time of contact can be prevented.

  Even if the objective optical system 3 is configured to be movable in a direction crossing the optical axis direction, the position of the objective optical system 3 is fixed to the microscope 2 by a simple method as necessary. As a result, it is possible to prevent inconveniences such as the objective optical system 3 moving after the optical axis is adjusted and the optical axis to be shifted, and to prevent blurring of the visual field during observation and obtain a more stable image. There are advantages.

In the above embodiment, the unit mounting portion 6 is movable in the optical axis direction of the objective optical system 3, but the stage 7 is movable in the optical axis direction of the objective optical system 3 instead. It may be.
Even if it does in this way, the effect similar to the said embodiment can be acquired.
In the above embodiment, the upright microscope 2 is used, but an inverted microscope may be used instead.

In the above embodiment, the microscope mounting portion 9 is mounted on the microscope 2 with the same structure as the mount of the objective optical system 3 by the microscope mounting screw 9a, but instead, it is the same as the mount of the revolver. It is good also as attaching to the microscope 2 with the structure of.
In the above-described embodiment, the objective attaching unit 11 may be attached with a revolver instead of the objective optical system 3. In this way, a plurality of objective optical systems 3 having different specifications can be attached to the revolver attached to the objective attaching portion 11, and observation can be performed while easily changing the magnification and the like.

In the above embodiment, the angle between the objective optical system 3 and the stage 7 may be adjustable. In this case, the stage 7 may be tilted, and the optical system including the objective optical system 3 and the position adjustment unit 4 may be tilted.
The support unit 5 fixed to the head of the mouse A may be arranged with the longitudinal direction of the insertion hole 5c inclined with respect to the optical axis of the objective optical system 3. Therefore, by adjusting the angle between the stage 7 and the objective optical system 3 so that the longitudinal direction of the insertion hole 5c is along the optical axis direction of the objective optical system 3, the small-diameter tip 3a can be easily inserted from an appropriate direction. It can be inserted into the hole 5c.

  At this time, for example, in a state where the specimen is fixed to the stage 7, a rod-shaped member having the same length as the objective optical system 3 or an object having the same shape as the outer cylinder of the objective optical system 3 is inserted into the insertion hole. The angle between the stage 7 and the objective optical system 3 is adjusted so that these central axes coincide with the optical axis of the microscope 2. Thereby, the angle between the stage 7 and the objective optical system 3 can be easily adjusted. Alternatively, the stage 7 and the objective optical system 3 can also be selected by attaching a reflecting member that reflects light at a position that closes the insertion hole 5c, such as a mirror, and selecting an angle at which the luminance of the image observed with the microscope 2 is highest. Can be easily adjusted.

1 Objective optical system position adjustment device 2 Microscope (observation optical system)
DESCRIPTION OF SYMBOLS 3 Objective optical system 3a Thin diameter front-end | tip part 4 Position adjustment unit 5 Support unit 5a Thin diameter part 5b Large diameter part 5c Insertion hole 5d Window 5e Transparent member 5f Inner surface 6 Unit attachment part 7 Stage 8 Fixing tool 9 Microscope attachment part 9a Microscope attachment Screw 9b Shading tube 10 Intermediate part 10a Intermediate part hole 10b Notch 11 Objective mounting part (holding part)
11a Objective mounting screw hole 12a X-axis guide (moving mechanism, linear guide)
12b Y-axis guide (moving mechanism, linear guide)
13 Centering screw (Optical axis adjusting means)
13a Screw part 13b Conical part 13c Rotating screw 14a Microscope side centering screw hole (optical axis adjusting means)
14b Objective-side centering hole (optical axis adjustment means)
15a Fixing screw (regulation means)
15b Fixing screw hole (regulation means)
16 shim (regulatory means)
17 Connecting member (regulation means)
100 Observation device A Mouse (living body)

Claims (7)

A position adjusting unit disposed between an objective optical system having a small-diameter tip and an observation optical system for observing light condensed by the objective optical system;
A substantially cylindrical support unit that has one end fixed to a living body and supports the small-diameter tip portion in an insertable and removable manner;
Wherein the position adjustment unit, wherein comprises a holding portion for holding the objective optical system, and a moving mechanism for movably supporting the holding unit in a direction intersecting the optical axis direction of the objective optical system, the narrow-diameter end portion When the holder is inserted into and removed from the support unit, the holding unit smoothly moves in the direction intersecting the optical axis direction by the moving mechanism, thereby adjusting the position of the objective optical system with respect to the support unit. ,
Said support unit, the other end portion, the objective optical system position adjustment device having a tapered inner surface which progressively diameter increases toward the other end. The position adjustment unit includes an optical axis adjustment unit that adjusts the position of the optical axis of the objective optical system held by the holding unit so as to coincide with the position of the optical axis of the observation optical system. The objective optical system position adjustment apparatus of description. The objective optical system position adjustment apparatus according to claim 1 , wherein the position adjustment unit includes a restriction unit that restricts movement of the holding unit.   The objective optical system according to claim 1, wherein the moving mechanism includes two linear guides that guide the holding unit in directions perpendicular to each other in a plane substantially perpendicular to the optical axis direction of the objective optical system. Positioning device.   The objective optical system position adjusting device according to claim 4, wherein the linear guide includes a bearing mechanism.   The support unit has a small-diameter portion fixed to the living body, a large-diameter portion having a larger diameter than the small-diameter portion, and a gradually increasing diameter from the small-diameter portion toward the large-diameter portion. 6. The objective optical system position adjusting device according to claim 1, wherein the objective optical system position adjusting device is a substantially cylindrical unit having the tapered inner surface. An observation apparatus provided with the objective optical system position adjustment apparatus according to any one of claims 1 to 6 .

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